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Biochemical and structural characterization of cross-linked enzyme aggregates (CLEAs) of organic solvent tolerant protease


Citation

Mohd Razib, Muhammad Syafiq and Raja Abdul Rahman, Raja Noor Zaliha and Mohd Shariff, Fairolniza and Mohamad Ali, Mohd Shukuri (2020) Biochemical and structural characterization of cross-linked enzyme aggregates (CLEAs) of organic solvent tolerant protease. Catalysts, 10 (1). art. no. 55. pp. 1-16. ISSN 2073-4344

Abstract

Cross-linked enzyme aggregates (CLEAs) is an immobilization technique that can be used to customize enzymes under an optimized condition. Structural analysis on any enzyme treated with a CLEA remains elusive and has been less explored. In the present work, a method for preparing an organic solvent tolerant protease using a CLEA is disclosed and optimized for better biochemical properties, followed by an analysis of the structure of this CLEA-treated protease. The said organic solvent tolerant protease is a metalloprotease known as elastase strain K in which activity of the metalloprotease is measured by a biochemical interaction with azocasein. Results showed that when a glutaraldehyde of 0.02% (v/v) was used under a 2 h treatment, the amount of recovered activity in CLEA-elastase was highest. The recovered activity of CLEA-elastase and CLEA-elastase-SB (which was a CLEA co-aggregated with starch and bovine serum albumin (BSA)) were at an approximate 60% and 80%, respectively. The CLEA immobilization of elastase strain K allowed the stability of the enzyme to be enhanced at high temperature and at a broader pH. Both CLEA-elastase and CLEA-elastase-SB end-products were able to maintain up to 67% enzyme activity at 60 °C and exhibiting an enhanced stability within pH 5–9 with up to 90% recovering activity. By implementing a CLEA on the organic solvent tolerant protease, the characteristics of the organic solvent tolerant were preserved and enhanced with the presence of 25% (v/v) acetonitrile, ethanol, and benzene at 165%, 173%, and 153% relative activity. Structural analysis through SEM and dynamic light scattering (DLS) showed that CLEA-elastase had a random aggregate morphology with an average diameter of 1497 nm.


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Additional Metadata

Item Type: Article
Divisions: Faculty of Biotechnology and Biomolecular Sciences
DOI Number: https://doi.org/10.3390/catal10010055
Publisher: MDPI
Keywords: CLEA; Solvent-tolerant protease; Glutaraldehyde; Immobilization
Depositing User: Nabilah Mustapa
Date Deposited: 03 May 2020 23:05
Last Modified: 03 May 2020 23:05
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.3390/catal10010055
URI: http://psasir.upm.edu.my/id/eprint/38188
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